Simultaneous isolation of the yeast cytosol and well-preserved mitochondria with negligible contamination by vacuolar proteinases

Disruption of yeast spheroplasts by DEAE-dextran in isoosmotic conditions allows isolation of relatively undamaged subcellular fractions from yeast. The preservation of mitochondria and vacuoles permits the simultaneous isolation of the cytosol with negligible contamination by vacuolar proteinases a...

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Bibliographic Details
Published in:FEBS letters 1983-06, Vol.156 (2), p.274-280
Main Authors: Schwencke, Jaime, Canut, Hervé, Flores, Alberto
Format: Article
Language:English
Subjects:
Cell Fractionation - methods
Cytosol - enzymology
Endopeptidases - isolation & purification
Glutamate Dehydrogenase - isolation & purification
Mitochondria - enzymology
Organoids - enzymology
Saccharomyces cerevisiae - enzymology
Saccharomyces cerevisiae - ultrastructure
Spheroplasts - enzymology
Temperature
Vacuoles - enzymology
Yeast Proteinase Vacuole Cytoplasm Proteolysis Mitochondria
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Summary:Disruption of yeast spheroplasts by DEAE-dextran in isoosmotic conditions allows isolation of relatively undamaged subcellular fractions from yeast. The preservation of mitochondria and vacuoles permits the simultaneous isolation of the cytosol with negligible contamination by vacuolar proteinases and therefore, virtually eliminates proteolytic artefacts.
ISSN:0014-5793
1873-3468
DOI:10.1016/0014-5793(83)80512-2